Method for monitoring molecular species within a medium

Abstract

Apparatus and methods for monitoring, analyzing, and / or discriminating molecular species, preferably a biomolecule, within a medium using a multisensor array (MSA) and multivariate processing. Biological compounds such as nucleotides and polynucleotides can be detected and analyzed. A reaction process such as an accumulation cycle of nucleic acids can be monitored, analyzed, and controlled using a multisensor array (MSA) and multivariate processing. Monitoring a biomolecule includes interrogating the medium, and preferably its gas phase, by coupling a sensor responsive to any changes of the medium and or biomolecule and its secondary products when, for example, an amplification reaction is proceeded. It is also a scope of the present invention to use direct detection and monitoring of biomolecular reactions in real-time without radioactive or fluorescent labeling. A preferred application is real-time polymerase chain reaction (PCR) detection.

Description

FIELD OF THE INVENTION[0001]The invention relates to an apparatus having a combination of multiple sensing probes or at least one multiple sensing probe and multivariate methods for monitoring and / or discriminating between molecular species, preferably biomolecules, within a medium preferably during a reaction process such as an accumulation cycle of nucleic acids.BACKGROUND OF THE INVENTION[0002]Several sensor technology developments have been described in the art. In particular, arrays of semiconductor sensors having sensitive and chemically-diverse interface materials capable of interacting with analytes of complex mixtures have previously been used. These can incorporate many operating, principles including: doped tin-oxide gas sensors, doped conductive polymers, field effect transistor (FET) devices, and optical fiber devices.[0003]Some sensors are based on a more specific chemical adsorption. For example, enzymes or antibodies can provide a more selective response when incorpo...

AI Technical Summary

Benefits of technology

[0031]With respect to PCR applications, the present invention obviates the exclusive requirement of attaching a fluorescent probe by preferably detecting any volatile substance by using at least one chemical sensor. Either an intercalator-based probe (e.g., ethidium bromide) where a chemical is intercalated within the double helix, or any of a component or secondary product from the master mix, buffer, primer or DNA molecule itself can be detected. This approach also provides a means of determining the effect of different reaction conditions on the efficacy of the amplification and so can provide insight into fundamentals and quality control of PCR processes.

Problems solved by technology

Furthermore, these examples have large complex fiber optic bundles.

This is important due to fouling of the sensors surfaces.

One particular problem with the above-described system involves the photobleaching of the dye molecules.

This limits the reliability and reproducibility of the sensor system.

These two methods lack the power to handle data with similar spectral features.

For example, there is little art on joining multivariate analysis with monitoring of any PCR process.

DNA and similar large biomolecules are hard to detect in gas phase.

These large biomolecules are hard to volatilize and are subject to degradation.

A major problem in analyzing genes is that they are rare targets in a complex genome that in mammals may contain as many as 100,000 genes.

Molecular genetics techniques currently used to overcome this problem are very time-consuming, involving cloning and methods for detecting specific DNA sequences.

Biotechnology NY, September, 11(9), 1026-30 (1993)] but one important drawback is the requirement for target-specific fluorogenic probes.

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